Team:ETHZ Basel/Biology/Status

From 2010.igem.org

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(testing of che-fusions)
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== testing of couplers ==
== testing of couplers ==
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We're planning to use the setup as for the testing of the localizers. In this case we're going to use the following biobricks:
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We're planning to use the setup as for the testing of the localizers.
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*localizer-PhyB and PIF3-cyFP or
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*tested biobricks: localizer-PhyB and PIF3-cyFP, localizer-PIF3 and PhyB-cyFP
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*localizer-PIF3 and PhyB-cyFP
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Images should look as in testing of localizer-fusions. Preliminary clarificaiton of the suitability of the approach are to be done.
Images should look as in testing of localizer-fusions. Preliminary clarificaiton of the suitability of the approach are to be done.

Revision as of 15:11, 6 August 2010

generation of subparts

subparts
Protein primers PCR product library of transformants single colonies storage vector insert
che proteins CheY-A recieved purified on stock on agar plate not available purified on stock on stock
CheY-C recieved purified in stock on agar plate not available purified on stock on stock
CheB-A recieved purified in stock on agar plate 3 on agar plate purified on stock on stock
CheB-C recieved purified in stock on agar plate 3 on agar plate purified on stock on stock
CheR-A recieved purified in stock on agar plate 3 on agar plate digest has to be redone
CheR-C recieved purified in stock on agar plate 3 on agar plate digest has to be redone
localizers TrigF-A recieved purified in stock on agar plate 3 on agar plate purified on stock
MreB-C recieved purified in stock in process
tetR-A recieved purified in stock on agar plate 3 on agar plate purified on stock on stock
couplers PIF3-A recieved purified in stock on agar plate 3 on agar plate purified on stock
PIF3-C recieved purified in stock on agar plate 3 on agar plate purified on stock
PhyB-C recieved purified in stock on agar plate 3 on agar plate purified on stock
reporters cyFP-A recieved purified in stock on agar plate 3 on agar plate purified on stock on stock
cyFP-C recieved purified in stock on agar plate 3 on agar plate purified on stock


generation of biobricks

We'll talk to Fabian. Apparently he has a nice protocol to generate acceptor vectors and transform E. coli sequentially in a 96 well plate.

testing of biobricks

testing of che-fusions

We're currently trying to establish an assay to test for the chemotactic behavour of E. coli. A first try with a "syringe test" failed. We'll redo the test and are looking for other possibilites. Maybe a swarm test?

  • tested biobricks: che-fusions
  • negative control: E. coli mutated for the accoding che-protein
  • positive control: wild type E. coli

testing of localicer-fusions

We are planning to use a fluorescence microscope in order to visualize the distribution of localizer-cyFP fusions within a single E. coli.

testing of couplers

We're planning to use the setup as for the testing of the localizers.

  • tested biobricks: localizer-PhyB and PIF3-cyFP, localizer-PIF3 and PhyB-cyFP

Images should look as in testing of localizer-fusions. Preliminary clarificaiton of the suitability of the approach are to be done.

bringing E. lemming alive

timetable

experimental plan